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Toxins 2015, 7(7), 2685-2700; doi:10.3390/toxins7072685

Biochemical Characterization of a Recombinant UDP-glucosyltransferase from Rice and Enzymatic Production of Deoxynivalenol-3-O-β-D-glucoside

1
Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, Vienna (BOKU), Konrad Lorenz Str. 24, 3430 Tulln, Austria
2
Christian Doppler Laboratory for Mycotoxin Metabolism and Center for Analytical Chemistry, Department of Agrobiotechnology (IFA-Tulln), BOKU, Konrad Lorenz Str. 20, 3430 Tulln, Austria
3
Department of Chemistry and Biochemistry, Mendel University in Brno, Zemědělská 1, 61300 Brno, Czech Republic
4
Biotechnology in Plant Production, Department IFA-Tulln, BOKU, Konrad Lorenz Str. 20, 3430 Tulln, Austria
5
Department of Biochemistry, University of Wisconsin, 433 Babcock Dr., Madison, WI 53706, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Richard A. Manderville
Received: 11 June 2015 / Revised: 10 July 2015 / Accepted: 16 July 2015 / Published: 21 July 2015
(This article belongs to the Section Mycotoxins)
View Full-Text   |   Download PDF [731 KB, uploaded 21 July 2015]   |  

Abstract

Glycosylation is an important plant defense mechanism and conjugates of Fusarium mycotoxins often co-occur with their parent compounds in cereal-based food and feed. In case of deoxynivalenol (DON), deoxynivalenol-3-O-β-D-glucoside (D3G) is the most important masked mycotoxin. The toxicological significance of D3G is not yet fully understood so that it is crucial to obtain this compound in pure and sufficient quantities for toxicological risk assessment and for use as an analytical standard. The aim of this study was the biochemical characterization of a DON-inactivating UDP-glucosyltransferase from rice (OsUGT79) and to investigate its suitability for preparative D3G synthesis. Apparent Michaelis constants (Km) of recombinant OsUGT79 were 0.23 mM DON and 2.2 mM UDP-glucose. Substrate inhibition occurred at DON concentrations above 2 mM (Ki = 24 mM DON), and UDP strongly inhibited the enzyme. Cu2+ and Zn2+ (1 mM) inhibited the enzyme completely. Sucrose synthase AtSUS1 was employed to regenerate UDP-glucose during the glucosylation reaction. With this approach, optimal conversion rates can be obtained at limited concentrations of the costly co-factor UDP-glucose. D3G can now be synthesized in sufficient quantity and purity. Similar strategies may be of interest to produce β-glucosides of other toxins. View Full-Text
Keywords: masked mycotoxin; glycosylation; sucrose synthase; UDP-glucose recycling; Fusarium masked mycotoxin; glycosylation; sucrose synthase; UDP-glucose recycling; Fusarium
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Michlmayr, H.; Malachová, A.; Varga, E.; Kleinová, J.; Lemmens, M.; Newmister, S.; Rayment, I.; Berthiller, F.; Adam, G. Biochemical Characterization of a Recombinant UDP-glucosyltransferase from Rice and Enzymatic Production of Deoxynivalenol-3-O-β-D-glucoside. Toxins 2015, 7, 2685-2700.

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